Cloning and expression of non-structural 3ABC gene from FMD virus serotype O in Escherichia coli

After about one hundred year after its identification, Foot and Mouth Disease (FMD) is the first viral disease in animals that impose a major risk to the world's livestock industry. The main method to identify animals vaccinated from animals infected with FMD is using of non-structural 3ABC protein as antigen in the ELISA kit. Hence, 3ABC gene of FMDV serotype O was isolated with PCR using specific primers containing BamHI and HinDIII restriction sites. The isolated fragment was then inserted in pTZ57R/T vector for cloning and nucleotide characterization. In order to produce recombinant antigens, 3ABC was inserted in pET21a (+) vector and then transform to E. coli BL21 (DE3). Recombinant protein production was induced with 1.5 mM IPTG. Production of recombinant proteins confirmed with SDS electrophoresis and Western blotting. The molecular weight of recombinant protein was determined approximately 50 kDa. The results showed that the produced protein can be used as an antigen in the ELISA kit for animals.